Histological and Immunohistochemical Characterization of Equine Anovulatory Haemorrhagic Follicles (AHFs)

Anovulatory haemorrhagic follicles (AHFs) are often the reason for ovulation failure in the mare. As the underlying factors that lead to AHF development are not well understood, it was of interest to investigate the vascularization of AHFs compared with normal follicles and corpora lutea (controls). In the present study, the ovarian cell populations investigated immunohistochemically included granulosa and luteal cells as well as various vascular structures. None of these cell types showed differences in the expression of vascular endothelial growth factor A (VEGF-A) between control ovaries containing normal follicles and corpora lutea and ovaries with AHFs. In contrast, a considerable reduction in the proportion of Flk-1-expressing cells, together with a decreased intensity of staining, was apparent in the AHFs. This greatly reduced expression of Flk-1 in the luteinized cells and the vascular structures of AHFs may lead to a distinct decrease in the potential pro-angiogenic activity of VEGF-A in these structures compared with the situation in normal follicles and corpora lutea. Furthermore, the authors suspect that the distinct expression of angiopoietin2 and VEGF-A seen in the cells within the inner fibrous layers of the AHFs was caused by hypoxia resulting from deficient vascularization, as suggested by the irregularity of the capillaries present in the luteinized wall of the AHF. In addition, whereas LH-receptor (LH-R) expression occurred uniformly in all stages of development of the corpora lutea in normal control ovaries, there was highly variable labelling for LH-R in all the AHFs examined, thereby indicating a possible numerical deficiency of LH-receptors in AHFs. The authors concluded that, despite the apparent expression of sufficient VEGF-A in the AHFs allows ovulation and corpus luteum formation, a relative lack of receptor, Flk-1, effects the pro-angiogenic activity of VEGF-A which could be a reason for ovulation failure associated with AHF formation.